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. 2015 Aug:482:51-9.
doi: 10.1016/j.virol.2015.03.024. Epub 2015 Mar 26.

Pathogenicity and pathogenesis of a United States porcine deltacoronavirus cell culture isolate in 5-day-old neonatal piglets

Qi Chen  1 , Phillip Gauger  1 , Molly Stafne  1 , Joseph Thomas  1 , Paulo Arruda  1 , Eric Burrough  1 , Darin Madson  1 , Joseph Brodie  1 , Drew Magstadt  1 , Rachel Derscheid  1 , Michael Welch  1 , Jianqiang Zhang  2
Affiliations

Pathogenicity and pathogenesis of a United States porcine deltacoronavirus cell culture isolate in 5-day-old neonatal piglets

Qi Chen et al. Virology. 2015 Aug.

Abstract

Porcine deltacoronavirus (PDCoV) was first identified in Hong Kong in 2009-2010 and reported in United States swine for the first time in February 2014. However, diagnostic tools other than polymerase chain reaction for PDCoV detection were lacking and Koch's postulates had not been fulfilled to confirm the pathogenic potential of PDCoV. In the present study, PDCoV peptide-specific rabbit antisera were developed and used in immunofluorescence and immunohistochemistry assays to assist PDCoV diagnostics. The pathogenicity and pathogenesis of PDCoV was investigated following orogastric inoculation of 5-day-old piglets with a plaque-purified PDCoV cell culture isolate (3 × 10(4) TCID50 per pig). The PDCoV-inoculated piglets developed mild to moderate diarrhea, shed increasing amount of virus in rectal swabs from 2 to 7 days post inoculation, and developed macroscopic and microscopic lesions in small intestines with viral antigen confirmed by immunohistochemistry staining. This study experimentally confirmed PDCoV pathogenicity and characterized PDCoV pathogenesis in neonatal piglets.

Keywords: Atrophic enteritis; Coronavirus; Immunohistochemistry; PDCoV; Peptide-specific antisera.

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Figures

Fig. 1
Fig. 1
Cytopathic effect (CPE) and IFA staining on PDCoV-infected ST cells or negative control ST cells (100× magnifications). (A) PDCoV CPE on ST cells at 24 h post infection; (E) negative control ST cells; (B and F) IFA staining with PDCoV M-peptide rabbit antiserum; (C and G) IFA staining with PDCoV N-peptide rabbit antiserum; (D and H) IFA staining with PDCoV S-peptide rabbit antiserum.
Fig. 2
Fig. 2
Evaluation of cross staining of PDCoV M-peptide rabbit antiserum, PEDV nucleocapsid monoclonal antibody conjugate, and TGEV polyclonal antibody conjugate on PDCoV-, PEDV-, TGEV-, and PRCV-infected cells.
Fig. 3
Fig. 3
Clinical assessment of PDCoV-inoculated pigs. (A) Watery diarrhea observed at 6 days post inoculation (DPI). (B) Average diarrhea scores. (C) Pigs at 5 DPI. (D) Average body weight.
Fig. 4
Fig. 4
Virus shedding in rectal swabs (A), sera (B) and various tissues (C) of PDCoV-inoculated pigs. Data labels (fractions) represent the number of PDCoV PCR positive pigs out of all pigs examined at that time point. The Ct values depicted in each figure were mean Ct values of PCR-positive pigs. The virus titers (Log10TCID50/ml) were mean virus titers of all pigs (both PCR-positive and negative pigs). Standard error bars are shown in each figure. At 3 DPI, sera were successfully collected from 8 out of 10 pigs.
Fig. 5
Fig. 5
Macroscopic and microscopic lesions and IHC staining. Thin-walled small intestines from a PDCoV-inoculated pig at 4 DPI (A). Hematoxylin and eosin-stained tissue sections of small intestine (middle jejunum) from a PDCoV-inoculated pig at 4 DPI (B, 600× magnification and C, 100× magnification). IHC-stained tissue sections of small intestine (middle jejunum) from a PDCoV-inoculated pig at 4 DPI (D, E, F with 20×, 100×, and 600× magnifications, respectively).
See this image and copyright information in PMC

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